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Offlinetubervegan
Stranger
Registered: 07/02/04
Posts: 76
Last seen: 2 years, 6 months
i'm losing hope, what would you do?
    #3346207 - 11/11/04 12:46 PM (12 years, 29 days ago)

Alright I just went through my 3rd attempt at growing and well its all fucked up again. Heres what I've done so far, first time I did PF tek and tried some weird shit and it totally didn't turn out and I shouldn't have expected to so forget about that one but then after that I tried PF tek and I got it to work except I only inoculated in one spot (I actually did two but one didn't turn out because I was trying to use a mycelium culture that I didn't realize was dead) and although things were going well and I had mycelium growth on one side, contaminants had enough time to form on the other.

Next I waited a while and tried again, this time with WBS I did two test jars because I was using some spores that had been sitting in water since the last time (approx. 6 months) one of these turned out and the other did not so I determined that one of the two spore solutions I had were still fine. Even the one that did turn out though did eventually grow a small contaminant.

So now I did a full load of jars and at first they seemed to be working out fine but in fact they are all contaminated, everyone of them.

Here is what I did, at first what I had been using to inoculate was a pipette because I was having problems finding long enough needles, this time I had them though. As a filter for the jars I used two coffee filters under the lid and one on top, but one those two test ones I used a layer of vermiculite.

For the sterilization, I let the jars all sit for about two days to let endospores germinate and then I autoclaved them all for 40 minutes each at about 17 PSI. I inoculated about half of them with new spore solution and when that ran out I started using the old stuff I had left over from before so about half of them are a combination. I inoculated each jar in 4 spots and flame sterilized the needles between each jar. To make the solution I sterilized a scalpel and a jar of water when cooled i opened the autoclave and scraped spores into the jar. The jar was covered with tin foil (sterile) and left outside the autoclave for about two days before being used. At all times there was an air ionizing running and the temp was 30 C +/- 3 degrees.

Another thing I should mention is that from leaving the cakes out for two days some of the seeds started to sprout and also from the autoclaving the jars seemed a little dry and some of the seed had cooked slightly from the high temps. Now after about 3 days I thought I saw growth I was excited, there were lines of mycelium forming where I had inoculated and not in between. I gave it some more time and then grew fast, the only thing I noticed is the mycelium looked a little dark and almost wet as opposed to the usual dry pure white look.

As time went on (its been just under 2 weeks now) It became evident that this was probably contamination. So today I took all the jars out and looked at everyone of them it turns out that almost all of them have this grey cobweb mold like substance growing exactly where I inoculated them and then some of them have light green mold in them (a few) a couple have sour rot, and some appear to have the beginnings of some lipstick mold in them?

meanwhile a few jars of reishi that I started a couple days after via grain to grain transfer are doing amazing, they are growing super fast and the jars (same batch as the others) are not contaminated at all, if anything I thought these jars would contaminate because I had to open them to the air..

anyways it seems that my spore water was contaminated right? but also some of them have other molds and bacteria growing in spots that don't correlate with the inoculation spots.

What advice can you give, I'm going to try it one more time and here is what I was thinking of doing: get a hepa filter, autoclave the jars and then boil them as well (I don't have a pressure cooker and I don't think I have money for one either), use a different spore print, I might actually get a pre-made spore syringe but I really don't have the money to be spending on stuff like that, and also because of money I'll probably go back to PF tek cause I already have the materials.

I can't think of anything else, what else can I do? has anyone ever seen this odd grey wet looking growth (I should note that as it grows denser it appears lighter)? I'll try to post pictures but I don't own a digital camera.

thanks and sorry for the long post.


Edited by tubervegan (11/11/04 01:27 PM)


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OfflineSilven
I trip,therefore weare.

Registered: 08/30/04
Posts: 2,071
Loc: El Mexicano
Last seen: 10 years, 8 months
Re: i'm losing hope, what would you do? [Re: tubervegan]
    #3346250 - 11/11/04 01:01 PM (12 years, 29 days ago)

1) Use paragraphs next time :wink:

2) Get a spore syringe, your sterility techniques are obviously not cutting it.

3) Don't use coffee filters

4) Follow the PF-Tek to a T

I can't give much more advice because it's too hard to pinpoint parts in your huge block of text without re-reading the whole thing. =P

This is why everyone suggests following the PF-Tek for your first time because it can get overwhelming trying to do everything right without having a step-by-step guide to reference from.

Use PF-Tek for Simple minds, that's how I started and it has not done me wrong yet.  Have only lost 3 jars to contams out of.. heh 48 PF-Tek jars.

Now I'm going to start grain though so it's a whole new ball game =)


--------------------
Born: 10/31/83, which makes me a Scorpio.

1) Scorpios are the most highly sexed of all the signs of the zodiac.
2) Scorpios are prone to excesses: booze, drugs, sex, bad puns, etc.
They usually exploit the weaknesses of others, who fall victim to
their capacity for total lust & sexual abberation.
3) Scorpios possess great intellectual curiosity & creative talent. They
think they are rebels & are arrogant, proud, conceited, and worth every
penny of it.

What do you bring to the table?


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OfflineKaptKid
Spaced Pirate
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Registered: 12/11/03
Posts: 6,247
Loc: Bright Side of the Sun
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Re: i'm losing hope, what would you do? [Re: Silven]
    #3346283 - 11/11/04 01:15 PM (12 years, 29 days ago)

Silven showed you a good course to follow but you got to steer that course.Its a proven course but its up to you to stay on course.Of course.hehe

As far as the contam's go, be sterial.Sounds like you got a contam problem to beat. GL

KK


--------------------
Child of the 60's, Tripping ever since.


:sun:


Edited by KaptKid (11/11/04 01:18 PM)


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Offlinetubervegan
Stranger
Registered: 07/02/04
Posts: 76
Last seen: 2 years, 6 months
Re: i'm losing hope, what would you do? [Re: Silven]
    #3346330 - 11/11/04 01:32 PM (12 years, 29 days ago)

Quote:

Silven said:
12) Get a spore syringe, your sterility techniques are obviously not cutting it.





whats throwing me off though is that I did the test jars and those did not have this problem, so although the contaminated started exactly where the jars were inoculated this didn't happen on the test jars which if anything were done in a less sterile manner, although the conditions were pretty much the same.

although I have made some minor adjustments to the PF tek I feel that if I try it I'm most likely going to end up with the same results, I can't figure out at which stage I'm getting contaminated...


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OfflineZeroArmy27
I didn't go to work for a month.
 Arcade Champion: Asteroids 2000

Registered: 09/01/04
Posts: 1,169
Loc: Middle of nowhere
Last seen: 6 years, 1 month
Re: i'm losing hope, what would you do? [Re: tubervegan]
    #3346410 - 11/11/04 01:55 PM (12 years, 29 days ago)

his time with WBS I did two test jars because I was using some spores that had been sitting in water since the last time (approx. 6 months)
-I found where your contamination came from.

:smile:

I don't use old spore solution. You're begging for contamination. Contams had 6 months to invade and thrive amongst your spore solution. :frown: not good.

Follow the Tek word for word your first time. You'll get a feel for what's going on. Don't try anything crazy or unusual until you are comfortable with growing and have some experience under your belt, and be smart about doing it too. Why would you experiment when you haven't even gotten results yet?


--------------------
"a monkey would fuck you up if you tried to put it in a autoclave" - Psychoslut

"it's not like the admins and mods are a tight-knit group of hippies that spend their life together in a log cabin tie-dying shirts and stringing beads inbetween bonghits." - Wiccan_Seeker


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Offlinetubervegan
Stranger
Registered: 07/02/04
Posts: 76
Last seen: 2 years, 6 months
Re: i'm losing hope, what would you do? [Re: ZeroArmy27]
    #3346499 - 11/11/04 02:16 PM (12 years, 29 days ago)

yes I understand how that might be the cause but, It worked in the test run, and then for the actual jars, I made new spore water which was used on half of them and still just the same they are contaminated so I'm hessitant to blame the spore solution even though it might seem like that.


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Offlinedoc34
Fungitarian
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Registered: 02/14/04
Posts: 2,659
Loc: Myceliaville !!!
Last seen: 1 month, 17 days
Re: i'm losing hope, what would you do? [Re: tubervegan]
    #3346769 - 11/11/04 03:40 PM (12 years, 29 days ago)

Try this VVVV

http://www.shroomery.org/index/par/26004

But you have to have a sterile syringe and practice sterility everywhere.


--------------------


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